Calcium in dark-adapted toad rods: evidence for pooling and cyclic-guanosine-3'-5'-monophosphate-dependent release. 1987

G L Fain, and W H Schröder
Department of Ophthalmology, Jules Stein Eye Institute, U.C.L.A. School of Medicine 90024.

1. We have used laser micromass analysis (l.a.m.m.a.) to investigate Ca uptake and release in intact 'red' rod photoreceptors in the dark-adapted retina of the toad, Bufo marinus. 2. With l.a.m.m.a. it is possible to measure separately the concentrations of each of the Ca isotopes. Rods normally containing almost exclusively 40Ca can be incubated in Ringer solution containing the stable isotopes 42Ca or 44Ca. In this way, the movements of Ca into and out of the rod can be separately determined. 3. When rods are incubated in darkness in high 44Ca (up to 20 mM), large amounts of 44Ca accumulate in the outer segment at a rate which increases with increasing external 44Ca concentration. However, this 44Ca appears not to exchange with the 40Ca originally present within the rod. This result suggests that the 40Ca may be sequestered within a pool which normally exchanges slowly with external Ca. 4. We explored Ca exchange in high-Ca solutions in more detail with double-isotope labelling. In these experiments, rods were first pre-loaded with Ca of one isotope (42Ca) and then incubated in Ringer solution containing another (44Ca). We could then measure separately the rate of exchange of the pre-loaded 42Ca with the 44Ca in the Ringer solution and with the 40Ca originally present within the rod in the sequestered pool. 5. These experiments show that the pre-loaded-Ca exchanges rapidly with Ca in the Ringer solution, at least in part by Ca-Ca exchange, but much more slowly with the Ca originally present within the rod. Thus Ca in the outer segments can exist in (at least) two pools: one which exchanges rapidly across the plasma membrane and is probably Ca free or loosely bound within the cytosol, and another which exchanges slowly and is probably Ca within the disks. 6. Although Ca sequestered within the outer segment normally exchanges quite slowly, it can be rapidly released if the extracellular free Ca is buffered to low levels with EGTA. The rate-limiting step for Ca release under these conditions appears not to be Na-Ca exchange, since the rate of Ca efflux is unchanged if the Na in the Ringer solution is substituted with choline. 7. Ca can also be released from the sequestered pool if rods are incubated in Ringer solution containing 100 or 500 microM-IBMX (3-isobutyl-1-methylxanthine).(ABSTRACT TRUNCATED AT 400 WORDS)

UI MeSH Term Description Entries
D010786 Photoreceptor Cells Specialized cells that detect and transduce light. They are classified into two types based on their light reception structure, the ciliary photoreceptors and the rhabdomeric photoreceptors with MICROVILLI. Ciliary photoreceptor cells use OPSINS that activate a PHOSPHODIESTERASE phosphodiesterase cascade. Rhabdomeric photoreceptor cells use opsins that activate a PHOSPHOLIPASE C cascade. Ciliary Photoreceptor Cells,Ciliary Photoreceptors,Rhabdomeric Photoreceptor Cells,Rhabdomeric Photoreceptors,Cell, Ciliary Photoreceptor,Cell, Photoreceptor,Cell, Rhabdomeric Photoreceptor,Cells, Ciliary Photoreceptor,Cells, Photoreceptor,Cells, Rhabdomeric Photoreceptor,Ciliary Photoreceptor,Ciliary Photoreceptor Cell,Photoreceptor Cell,Photoreceptor Cell, Ciliary,Photoreceptor Cell, Rhabdomeric,Photoreceptor Cells, Ciliary,Photoreceptor Cells, Rhabdomeric,Photoreceptor, Ciliary,Photoreceptor, Rhabdomeric,Photoreceptors, Ciliary,Photoreceptors, Rhabdomeric,Rhabdomeric Photoreceptor,Rhabdomeric Photoreceptor Cell
D012160 Retina The ten-layered nervous tissue membrane of the eye. It is continuous with the OPTIC NERVE and receives images of external objects and transmits visual impulses to the brain. Its outer surface is in contact with the CHOROID and the inner surface with the VITREOUS BODY. The outer-most layer is pigmented, whereas the inner nine layers are transparent. Ora Serrata
D002024 Bufo marinus A species of the true toads, Bufonidae, becoming fairly common in the southern United States and almost pantropical. The secretions from the skin glands of this species are very toxic to animals. Rhinella marina,Toad, Giant,Toad, Marine,Giant Toad,Giant Toads,Marine Toad,Marine Toads,Toads, Giant,Toads, Marine
D002118 Calcium A basic element found in nearly all tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes. Coagulation Factor IV,Factor IV,Blood Coagulation Factor IV,Calcium-40,Calcium 40,Factor IV, Coagulation
D003623 Dark Adaptation Adjustment of the eyes under conditions of low light. The sensitivity of the eye to light is increased during dark adaptation. Scotopic Adaptation,Adaptation, Dark,Adaptation, Scotopic
D006152 Cyclic GMP Guanosine cyclic 3',5'-(hydrogen phosphate). A guanine nucleotide containing one phosphate group which is esterified to the sugar moiety in both the 3'- and 5'-positions. It is a cellular regulatory agent and has been described as a second messenger. Its levels increase in response to a variety of hormones, including acetylcholine, insulin, and oxytocin and it has been found to activate specific protein kinases. (From Merck Index, 11th ed) Guanosine Cyclic 3',5'-Monophosphate,Guanosine Cyclic 3,5 Monophosphate,Guanosine Cyclic Monophosphate,Guanosine Cyclic-3',5'-Monophosphate,3',5'-Monophosphate, Guanosine Cyclic,Cyclic 3',5'-Monophosphate, Guanosine,Cyclic Monophosphate, Guanosine,Cyclic-3',5'-Monophosphate, Guanosine,GMP, Cyclic,Guanosine Cyclic 3',5' Monophosphate,Monophosphate, Guanosine Cyclic
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D013997 Time Factors Elements of limited time intervals, contributing to particular results or situations. Time Series,Factor, Time,Time Factor
D015056 1-Methyl-3-isobutylxanthine A potent cyclic nucleotide phosphodiesterase inhibitor; due to this action, the compound increases cyclic AMP and cyclic GMP in tissue and thereby activates CYCLIC NUCLEOTIDE-REGULATED PROTEIN KINASES 3-Isobutyl-1-methylxanthine,Isobutyltheophylline,IBMX,1 Methyl 3 isobutylxanthine,3 Isobutyl 1 methylxanthine
D066298 In Vitro Techniques Methods to study reactions or processes taking place in an artificial environment outside the living organism. In Vitro Test,In Vitro Testing,In Vitro Tests,In Vitro as Topic,In Vitro,In Vitro Technique,In Vitro Testings,Technique, In Vitro,Techniques, In Vitro,Test, In Vitro,Testing, In Vitro,Testings, In Vitro,Tests, In Vitro,Vitro Testing, In

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